N-aminoxy-acetyl-n&#39;-isonicotinoyl-hydrazine and the pharmaceutically acceptable salts thereof

ABSTRACT

N,N&#39;&#39;-DIACYL HYDRAZINE DERIVATIVES HAVING TUBERCULOSTATIC ACTIVITY AND CORRESPONDING TO THE GENERAL FORMULA   X-NH-O-CH(-R)-CO-NH-NH-AC   WHEREIN X REPRESENTS HYDROGEN OR AN ACYL RADICAL, R REPRESENTS HYDROGEN OR A SUBSTITUTED OR UNSUBSTITUTED ALKYL, ARALKYL OR ARYL GROUP, AND AC IS THE ACYL RADICAL OF A SUBSTITUTED OR UNSUBSTITUTED ALIPHATIC, AROMATIC OR HETEROCYCLIC CARBOXYLIC ACID, AS WELL AS THE PHARMACEUTICALLY ACCEPTABLE ACID ADDITION SALTS THEREOF AND/OR THE OPTICALLY ACTIVE ISOMERS OF SUCH COMPOUND CONTAINING AN ASYMMERTIC CARBON ATOM.

United States Patent N AMINOXY-ACETYL N ISONICOTINOYL- HYDRAZINE AND THEPHARMACEUTICALLY ACCEPTABLE SALTS THEREOF Lajos Kisfaludy, Agnes Patthy,nee Lukats, Lajos Dancsi, Gyorgy Fekete, and Istvan Szabo, Budapest,Hungary, assignors to Richter Gedeon Vegyeszeti Gyar Rt, Budapest,Hungary N Drawing. Filed Dec. 28, 1970, Ser. No. 102,226

Int. Cl. C07d 31/44 US. Cl. 260-295 AM 1 Claim ABSTRACT OF THEDISCLOSURE N,N'-diacyl hydrazine derivatives having tuberculostaticactivity and corresponding to the general formula wherein X representshydrogen or an acyl radical, R represents hydrogen or a substituted orunsubstituted alkyl, aralkyl or aryl group, and Ac is the acyl radicalof a substituted or unsubstituted aliphatic, aromatic or heterocycliccarboxylic acid, as well as the pharmaceutically acceptable acidaddition salts thereof and/or the optically active isomers of suchcompound containing an asymmetric carbon atom.

The invention relates to the preparation of novel N, N'-diaoyl-hydrazinederivatives of tuberculostatic activity.

Among the tuberculostatically active compounds, a very important placeis taken by certain hydrazides, in view of the fact that theisonicotinic acid hydrazide is considered up to now as the most potentantituberculotic agent. On the other hand, the bacteriostatic effect ofaminoXy-acetic acid is known for a long time [cf. C. B. Favour,Bacteriol. 55, 1 (1948)]; the eifect of this compound was examined inmore detail, however, only since it has been discovered that thecycloserine, which contains the aminoxy group closed into a ring, is animportant drug in the therapy of human tuberculosis [cf. F. A. Kuel,etc. I. Am. Chem. Soc. 77, 2344 (1955); P. H. Hidy, etc., I. Am. Chem.Soc,. 77, 2346 (1955)]. Numerous further open-chain aminoxy compoundshave been described later [cf. McHale, etc., J. Chem. Soc. (1960), 225;P. Mamalis, etc., J. Chem. Soc. (1960), 229; A. Frank, K. Riede,Monatshefte Chem. 92, 725 (1961); E. Tesla etc., Helv. Chim. Acta 46,766 (1963); P. Mamalis, etc. J. Med. Chem. 6, 684 (1965); V. Markovaetc.; Khim. Farm. Zh. 3, 13 (1969)], and their effects on variousmicro-organisms have been investigated [cf. S. A. Price, etc., Brit. J.Pharm. 15, 243 (1960)], but no compound having a potent activity againstMycobacterium tuberculosis could be found in this family of aminoxyderivatives.

It has been found now surprisingly that the simple u-aminoxy-carboxylicacid hydrazides not described in the literature hitherto andcharacterized by the general Formula I wherein X represents hydrogen oran acyl radical, R represents hydrogen or a substituted or unsubstitutedallcyl, aralkyl or aryl group, and Ac is the acyl radical of asubstituted or unsubstituted aliphatic, aromatic or heterocycliccarboxylic acid, as well as the therapeutically acceptable acid additionsalts thereof and/or the optically active isomers of such compoundscontaining an asymmetric carbon atom, have very favourabletuberculostatic etfects.

ICC

In the compounds of the general Formula I according to the presentinvention X represents mostly hydrogen, it may, however, represent alsoan acyl radical, possibly containing also a group capable to form salts.

The compounds containing hydrogen in the place of R are aminoxy-aceticacid hydrazides, whereas those in which R is different from hydrogen,are ot-aminoxy-carboxylic acid hydrazides containing an asymmetricalcarbon atom and can thus exist in optically active forms. The symbol Acrepresents in accordance with the above definition mainly the acylradical of a heterocyclic carboxylic acid, but it may represent the acylradical e.g. of an amino acid or an amino-aryl carboxylic acid too.

The new compounds of the general Formula I of the invention may beprepared in various ways. So e.g. an aaminoxy-carboxylic acid derivativeof the general Formula II wherein A represents an acyl radical, or if inthe desired reaction product X represents hydrogen, then A can be aprotective group apt for the temporary protection of the amino group, ase.g. a benzyloxy-carbonyl or tert.- butyloxycarbonyl group, R has thesame meaning as above, and B represents a hydroXyl group or a groupcapable to activate the carboxyl group, e.g. a pentachlorophenoxy group,a halogen atom or an azido group, is reacted with a hydrazide of thegeneral Formula III wherein Ac has the same meaning as above.

According to another method, an a-arninoxycarboxylic acid hydrazide ofthe general Formula IV A-NHo-(lJH-o0NH-NHa wherein A and R have the samemeanings as above, is

reacted with a carboxylic acid derivative of the general Formula V AcBwherein Ac and B have the same meaning as above.

The compound of the general Formula VI obtained by the above methodswherein A, R and Ac have the same meanings as above, may be converted bysplitting off the protective group A in a per se known manner andreplacing it by the desired acyl group X into the end product of thegeneral Formula I or into a therapeutically acceptable acid additionsalt thereof. If the acyl group Ac is the acyl radical of a carboxylicacid, containing also itself a group to be temporarily protected, e.g.an amino group, this may be protected preferably by the same protectivegroup A which has been used to protect the amino group of the startingcompound, in order to ensure the uniform progress of the reaction.

The compounds of the general Formula I obtained by the methods describedabove from racemic starting compounds of the Formula H or IV,respectively, may be resolved by per se known methods into the opticalenantiomers.

In preparing the compounds of the general Formula VI, either by reactinga compound of the Formula II with one of the Formula III, or by reactinga compound of the Formula IV with one of the Formula V, the startingmaterials are reacted in an organic solvent, preferably indimethylformamide, at room temperature.

The reaction mixture can be easily worked up, because after removing thesolvent from the reaction mixture the desired product can becrystallized easily from the residual mixture by the aid of an organicsolvent or of a mixture of a water miscible organic solvent with water.

The compound of the general Formula VI is then treated-according to thenature of the protective groupwith hydrogen bromide in glacial aceticacid or with ethyl acetate saturated by hydrogen chloride, whereby theprotective group is split oif and simultaneously the corresponding saltof the end product of the Formula I is formed, which is insoluble inether and can be thereby easily precipitated from the reaction mixture.

According to an especially preferred way of performance of the process,tert.butyloxycarbonyl or carbobenzoxy protective groups are used for thetemporary protection of the amino group of the startinga-aminoxycarboxylic acid derivative, and the activation of the carboxylgroup is achieved by converting it into pentachlorophenyl ester. In thisWay the compound of the general Formula V1 is obtained with excellentyields. In case of standing in the place of Ac an acid radicalcontaining also a basic group-cg. if Ac is an isonicotinoyl group thepentachlorophenolate of the compound of the Formula VI is formed in thisway; this is a stable salt which can be isolated and purified easily andso the recovery of the compound is considerably facilitated.

The compound of the Formula VI can be converted in a per se known mannerby acid treatment into the end product of the general Formula I.Depending on the reaction conditions applied, the compounds of thegeneral Formula I are obtained in the form of free bases or of salts.From the salts obtained, the bases may be liberated in per se knownmanner, and the bases may be converted into acid addition salts, formedpreferably with therapeutically acceptable acids.

In experiments carried out in vitro, numerols compounds of the generalFormula I prepared according to the invention, inhibited considerablythe growth of the strain H R of Mycobacterium tuberculosis as well asthe growth of strains resistant to isonicotinic acid hydrazide,p-aminosalicylic acid and streptomycin. The extent of inhibition isespecially marked in the case of the compound described in Example 1,where the inhibiting concentration was lower than 0.1 mg./ml. The testsperformed in vivo on guinea-pigs and on white mice, confirmed the invitro established effectiveness of the said compound. The guinea-pigshave been infected subcutaneously by 0.01 mg. of bacteria (calculated onwet weight). The drug treatment has been commenced on the next day afterthe infection and has been performed by oral administration of thecompound in doses of 3 to 7 mg./ kg. during 90 days. When dissecting theanimals, the pathological lesions of five organs have been evaluated andrecorded by the-figures O to 5, according to the gravity of the lesion.The average value shown by the animals of the control group was 15.5,while the treated animals have shown an average value of 3.1. Thecompounds tested inhibited in doses of 7 mg./kg. virtually completelythe generalization of the infection by Mycobacteria. In the case of thetests performed on white mice, the grade of lesion of the lungs has beenrecorded after the necrotomy. The animals of the untreated control grouphave shown an average lung-index of 4.2, while the value obtained withthe animals treated by doses of 15 mg./kg. has been 0.4. A furthercontrol group treated with similar doses of viomycin has shown alung-index of 2.08. On the basis of the above experimental results, thetuberculostatic activity of the compound described in Example 1,(N-aminoxy-acetyl N isonicotinoyl-hydrazine) is of the same order asthat of the most potent antituberculotic drugs. To establish thetoxicity of the said compound in a preliminary experiment, guinea-pigswere fed for three months with doses of 15 mg./kg. daily, then theanimals were killed and histological section fixed with formol were madeof the livers, kidneys, lungs, heartmuscles and spleens, and these wereexamined after the usual hematoxylin-eosine staining. The histologicalexamination revealed no lesions.

For therapeutical purposes, the compounds of the general Formula I maybe administered orally and/or parenterally, in the form of tablets,drages, injections, infusions or suppositories. The daily doses foradult persons may be 7 to 50 mg./kg.

The preparation of the new compound is illustrated in more details bythe following examples. The R values given in the examples have beenestablished by thin layer chromatography on silica gel layer, in thesolvent system n-hexane-glacial acetic acid-chloroform 1111-8, thechromatograms have been developed with chlorine and toluidine. Thechemical structures of the compounds described in the examples have beenchecked occasionally also by IR or NMR spectroscopy.

EXAMPLE 1 (a) N-(N"-benzyloxycarbonylaminoxy-acetyl) -N'-isonicotinoylhydrazine (a 4.73 g. (10 mmol.) of N-benzyloxycarbonylaminoxy-aceticacid pentachlorophenyl ester are dissolved in 50 ml. of abs. dimethylformamide, 1.37 g. (10 mmol.) of isonicotinic acid hydrazide are addedthereto and the mixture is allowed to stand overnight. The solvent isthen distilled oil? under reduced pressure and the residue iscrystallized from ethanol. The pentachlorophenolate salt ofN-(N"-benzyloxycarbonyl-aminoxy-acetyl)-N'-isonico tinoyl hydrazine isobtained, yield: 5.37 g. (88%); M.P. l60-162 C.; R =0.18.

Analysis.C-alculated (percent): C, 43.3; H, 2.7; Cl, 29.1. Found(percent): C, 43.3; H, 2.8; CI, 29.1.

The salt prepared in this way is subjected to partition between 25 ml.of ethyl acetate and 15 ml. of N-hydrochloric acid, the aqueous layer isneutralized by adding NaI-ICO and then allowed to stand cold. Theprecipitated product is separated by filtration and recrystallized from50% ethanol. 2.64 g. (87%) of N-(N-benzyl-0xycarbonyl-aminoxy-acetyl)-N-isonicotinoyl-hydrazine are obtained; M.P.112113 C.; R;=0.18.

Analysis.-Calculated (percent): C, 55.8; H, 4.7; N, 16.3. Found(percent): C, 55.8; H, 4.9; N, 16.3.

(a 2.15 g. (9 mmol.) of N-benzyloxycarbonylaminoxy-acetic acid hydrazideare dissolved in 30 ml. of abs. dimethylformamide, 3.34 g. (9 mmol.) ofisonicotinic acid pentachlorophenyl ester are added thereto and thereaction mixture is allowed to stand overnight. The solvent is thendistilled off under reduced pressure and the residue is recrystallizedfrom ethanol. 3.62 g. (66%) of the pentachlorophenolate salt areobtained, the physical characteristics of the product are the same as inparagraph (a this salt can be converted intoN-(N"-benzyloxycarbonyl-aminoxy-acetyl)-N-isonicotinoyl hydrazine by themethod described in paragraph (a (a 2.25 g. (10 mmol.) ofN-benzyloxycarbonylaminoxy-acetic acid are dissolved in 20 ml. ofdimethyl formamide and 1.37 g. (10 mmol.) of isonicotinic acid hydrazideand then 2.06 g. (10 mmol.) of dicyclohexyl carbodiimide are added tothe stirred and cooled solution and the mixture is further stirred forone day at room temperature. The dicyclohexyl urea is filtered off andthe filtrate is evaporated to dryness under reduced pressure. Theresidue is crystallized from aqueous methanol. 2.26 g. (66%) ofN-(N"-benzyloxycarbonyl-aminoxy-acetyl)- N-isonicotinoyl hydrazine areobtained; the physical characteristics of the product are the same as inparagraph (a (b) N- (N"-tert.-butyloxycarbonyl-aminoxy-acetyl)-N'-isonicotinoyl hydrazine (b 4.39 g. 10 mmol.) ofNtert.-butyloxycarbonylaminoxy-acetic acid pentachlorophenyl ester aredissolved in 50 ml. of dimethylformarnide, 1.37 g. (10 mmol.) ofisonicotinic acid hydrazide are added thereto and the reaction mixtureis allowed to stand overnight. The solvent is then distilled off underreduced pressure, the residue is subjected to partition between 50 ml.of ethyl acetate and 30 ml. of N hydrochloric acid at C. The productprecipitated by neutralization of the aqueous layer is then extractedinto ethyl acetate, the solution is dried and the solvent is distilledolf. The residue is crystallized from ethyl acetate. 2.36 g. (76%) ofN-(N-tert.-butyloxycarbonyl-aminoxy-acetyl)-N-isonicotinoyl-hydrazineare obtained; M.P. 168169 C.; R =0.16.

Analysis.--Calculated (percent): C, 50.3; H, 5.9; N, 18.0. Found(percent): C, 50.1; H, 6.1; N, 18.1.

(b 1.91 g. mmol.) of N-tert.-butyloxy-carbonylaminoxy-acetic acid aredissolved in ml. of abs. dimethyl formamide, 1.37 g. (10 mmol.) ofisonicotinic acid hydrazide and 2.06 g. (10 mmol.) of dicyclohexylcarbodiimide are added to the stirred solution and the stirringis'continued further for 24 hours. After filtering oif the dicyclohexylurea precipitated, the filtrate is evaporated to dryness under reducedpressure and the residue is crystallized from ethyl acetate. 2.28 g.(61%) of N- (N"-tert.-butyloxycarbonyl-aminoxy-acetyl) Nisonicotinoyl-hydrazide are obtained, the physical characteristics ofthis product are the same as in paragraph (b (0)N-aminoxy-acetyl-N'-isonicotinoyl-hydrazinedihydrobromide 97.0 g. (159mmol.) of N-(N"-benzyloxy-carbonylaminoxy-acetyl)-N-isonicotinoylhydrazine pentachlorophenolate salt are suspended in 250 ml. of glacialacetic acid and 250 ml. of a 4 M solution of hydrobromic acid in glacialacetic acid are added thereto under stirring and excluding the airhumidity. After further stirring for one hour, 4.5 liters of dry etherare added to the reaction mixture. The precipitate is collected byfiltration, washed with ether, then dissolved in 750 ml. of abs.methanol and crystallized by adding 1 liter of ether. 58.0 g. (98%) ofthe di-hydrobromide salt of N-aminoxy-acetyl-N-isonicotinoyl-hydrazineare obtained; M.P.: l66l67 C.

Analysis.Calculated (percent): C, 25.8; H, 3.2; N, 15.1; Br, 43.0. Found(percent): C, 25.8; H, 3.3; N, 15.2; Br, 42.9.

(d) N-aminoxy-acetyl-N-isonicotinoyl-hydrazine dihydrochloride 1.6 g.(5.2 mmol.) of N-(N"-tert.-butyloxycarbonylaminoxy-acetyl) Nisonicotinoyl-hydrazine are suspended in 7.0 ml. of dry ethyl acetate,and 8.0 ml. of a 4 M solution of hydrogen chloride in ethyl acetate areadded at 20 C. to the stirred suspension. After stirring for further 30minutes 80 ml. of dry ether are added to the reaction mixture, theprecipitated product is collected by filtration, washed with ether andthen recrystallized from methanol. 1.28 g. (87%) of the di-hydrochloridesalt of N-aminoxy-acetyl N isonicotinoyl-hydrazine (87%) are obtained;M.P.: 182-l84 C.

Analysis.Calculated (percent): C, 33.9; H, 4.3; CI, 25.1. Found(percent): C, 34.0; H, 4.3; CI, 25.0.

EXAMPLE 2 (a) N,N'-di- (N"-benzyloxycarbonyl-aminoxy-acetyl) hydrazine2.36 g. (5 mmol.) of N-benzyloxycarbonyl-aminoxyacetic acidpentachlorophenyl ester are dissolved in 25 m1. of abs. dimethylformamide, 0.17 ml. (2.5 mmol.) of hydrazine hydrate are added to thestirred and cooled solution and the reaction mixture is allowed to standovernight. The solvent is then distilled off under reduced pressure andthe residue is treated with ether and crystallized from ethanol. 0.8 g.(72%) of N,N-di-(N"-benzyloxycarbonyl-aminoxy-acetyl) hydrazine areobtained, M.P. 164166 C.; R,=0.92.

Analysis.-Calculated (percent): C, 53.8; H, 5.0. Found (percent): C,53.7; H, 4.9.

(b) N,N'-di- (aminoxy-acetyl -hydr azine di-hydrobromide This compoundwas prepared from 3.2 g. of N,N'-di (N"-benzyloxycarbonyl-aminoxy-aceticacid hydrazide 'by the method described in Example 1(0). 2.3 g. (94%) ofN,N'-di-(aminoxy-acetyl) hydrazine di hydrobromide were obtained; M.P.178180 C. (from ethanol/ether).

Analysis.Calculated (percent): C, 14.2; H, 3.6; N, 16.5; Br, 46.7.'Found (percent): C, 14.2; H, 3.7; N, 16.4; Br, 46.8.

EXAMPLE 3 (a) N- (N"-benzyloxycarbonyl-glycyl -N-(N"'-benzyloxycarbonyl-aminoxy-acetyl) -hydrazine (a This compound wasprepared from 2.0 g. of N- benzyloxycarbonyl-glycine hydrazide and 3.78g. of N- benzyloxycarbonyl aminoxy acetic acid pentachlorophenyl ester,by the method described in Example 1(a 3.34 g. (97%) ofN-(N"-benzyloxycarbonyl-glycyl)-N'-(N"-benzyloxycarbonyl-aminoxy-acetyl)-hydrazine were obtained; M.P.148-149 C. (from ethanol); R =0.12.

Analysis.Calculated (percent): C, 55.8; H, 5.2. Found (percent): C,55.7; H, 5.3.

(a The same compound has been prepared also from 1.0 g. ofN-benzyloxycarbonyl-amino-acetic acid hydrazide and 1.74 g. ofN-benzyloxycarbonyl-glycine pentachlorophenyl ester by the methoddescribed in Example 1 (a 1.25 g. (72%) of the above product wereobtained, having the same physical characteristics as described inparagraph (a (b) N-glycyl-N'-aminoxy-acetyl-hydrazine di-hydrobromideThis compound was prepared from 1.05 g. of N-(N-benzyloxycarbonyl-glycyl)-N (N"'benzyloxycarbonylaminoxy-acetyl)-hydrazine by the method described inExample 1(0). 0.71 g. of N-glycyl-N'-(aminoxyacetyl)-hydrazinedi-hydrobromide was obtained; M.P. 179184 C. (from methanol/ether).

Analysis.Calculated (percent): C, 14.8; H, 3.8; Br, 49.4. Found(percent): C, 14.9; H, 3.9; Br, 49.2.

EXAMPLE 4 (a) N-(N"-benzyloxycarbonyl-L-alanyl)-N-(N-benzyloxycarbonyl-aminoxy-acetyl) -hydrazine This compound was preparedfrom 11.82 g. of N-benzyloxycarbonyl-aminoxy-acetic acidpentachlorophenyl ester and 6.4 g. of N-benzyloxycarbonyl-L-alaninehydrazide by the method described in Example 1 (a 7.92 g. (71%) ofN-(N"-benzyloxycarbonyl-L-alanyl)-N-(N"'benzyloxycarbonyl-aminoxy-acetyl) hydrazine were obtained, M.P. -186 C;R =0.7.

Analysis-Calculated (percent): C, 56.8; H, 5.4. Found (percent): C,56.9; H, 5.5.

(b) N-L-alanyl-N-aminoxy-acetyl-hydrazine di-hydrobromide This compoundwas prepared from 3.0 g. of N-(N"- benzyloxycarbonyl-L-alanyl) N (N"'benzyloxycarbonyl-aminoxy-acetyl)-hydrazine by the method described inExample 1(c). 2.1 g. (92%) of N-L-alanyl-N-aminoxy-acetyl-hydrazinedi-hydrobromide were obtained; M.P. 108-110 C. (from ethanol/ether).

Analysis.Calculated (percent): C, 17.7; H, 4.2; Br, 47.3. Found(percent): C, 17.8; H, 4.1; Br, 47.2.

EXAMPLE 5 (a) N- N '-tert.-butyloxycarbonyl-aminoxy-acetyl -N-(N"-tert.-butyloxycarb onyl-L-seryl) -hydrazine This compound wasprepared from 3.94 g. of N-tert.- butyloxycarbonyl-aminoxy-acetic acidpentachlorophenyl ester and 2.19 g. ofN-tert.-butyloxycarbonyl-L-seryl-hydrazine by the method described inExample 1(b 3.4 g. (87 of N-(N"-tert. butyloxycarbonylaminoxyacetyl)-N'-(N"'-tert.-butyloxycarbonyl L seryl) hydrazine wereobtained; M.P. 118 C. (from methanol); R =0.1.

Analysis.-Calcu1ated (percent): C, 45.9; H, 7.2; N, 14.3. Found(percent): C, 46.0; H, 7.4; N, 14.4.

(*b) N-L-seryl-N- aminoxy-acetyl hydrazine dihydrobromide This compoundwas prepared by treating 3.15 g. of the product described in Example5(a) with a 4 M solution of hydrobromic acid in trifiuoroacetic acid, bythe method described in Example 1(d). 1.98 g. (81%) of N-L-seryl-N'-(aminoxy-acetyl)-hydrazine di hydrobromide were obtained;M.P. 139141 C. (from ethanol/ether).

Analysis.-Calculated (percent): C, 20.0; H, 4.0; Br, 45.2. 'Found(percent): C, 20.1; H, 4.1; Er, 45.2.

EXAMPLE 6 (a) N-(N"-benzyloxycarbonyl-aminoxy-acetyl) -N'-(N-benzyloxycarbonyl-L-phenylalanyl)-hydrazine This compound was preparedfrom 4.25 g. of N-benzyloxycarbonyl-arnioxy-acetic acidpentachlorophenyl ester and 3.2 g. ofN-benzyloxycarbonyl-L-phenyl-alanyl-hydrazine, as described in Examplela 4.19 g. (81%) of N-(N benzyloxycarbonyl-aminooxy-acetyl) N (N"-benzyloxycarbonyl-L-phenylalanyl) hydrazine were obtained; M.P. 199-200C. (ethanol); R =0.35.

Analysis.-Calculated (percent): C, 62.3; H, 5.4. Found (percent): C,62.2; H, 5.4.

(b) N-aminoxy-acetyl-N'-L-phenylalanyl-hydrazine dihydrobromide Thiscompound was prepared from 3.0 g. of N-(N"-benzyloxycarbonyl-aminoxy-acetyl)-N'-(N'benzyloxycarbonyl-L-phenylalanyl)hydrazine, as described in Example ld).2.17 g. (90%) of N-aminoxy-acetyl-N-L- phenylalanyl-hydrazinedihydrobromide were obtained; M.P. 143 C.

Analysis.Calculated (percent): Br, 38.6. Found (percent): Br. 38.3.

EXAMPLE 7 (a) N- (N-tert.-butyloxycarb onyl-a-arninoxy-propionyl)N-isonicotinoyl-hydrazine pentachlorophenolate (b)N-(ot-aminoxvpropionyl)-N-isonicotinoylhydrazine-dihydrochloride 1.89 g.(3.2 mmol.) ofN-(N"-tert.-butyl-oxycarbonyla-aminoxy-propionyl)-N'-isonicotinoyl-hydrazinepentachlorophenolate are suspended in a 4 mol./liter solution ofhydrogen chloride in ethyl acetate and the reaction is stirred for 30minutes. 10 ml. of dry ether are then added to the solution, theprecipitate is separated by filtration, washed with ether recrystallizedfrom the mixture of anhydrous ethanol and ether. 0.86 g. (90%) ofN-(ocaminoxy-propionyl)-N'-is0nicotinoyl hydrazine dihydrochloride areobtained; M.P. 181 C.; [a] =+36.6 (c.=1, ethanol 96%).

Analysis.Calculated (percent): C, 36.4; H, 4.8; CI, 23.8. Found(percent): C, 36.5; H, 4.8; Cl, 23.8.

EXAMPLE 8 N-(w-aminoxy-fi-phenyl-propionyl) -N'-isonicotinoyl hydrazinedihydrochloride This compound was prepared from 2.4 g. (6 mmol.) of N(N"-tert.butyloxycarbonyl-a-aminoxy-fl-phenyl-propionyl)-N'-isonicotinoylhydrazine as described in Example 1(d). 1.85 g. (83%) ofN-(a-aminoxy-fl-phenyl-propionyl.)-N'-isonic0tinoyl hydrazinedihydrochloride were obtained; M.P. 92 C. (methanol/ether); [m] '+42.0(c.=1, ethanol).

Analysis.Calculated (percent): C, 48.3; H, 4.8; N, 15.0; C1, 19.0. Found(percent): C, 48.2; H, 5.0; N, 14.9; CI, 19.2.

EXAMPLE 9 N'-(N"-aminoxy-acetyl)-aminoxy-acetyl-N'-aminoxyacetylhydrazinedihydrochloride 2.22 g. (0.008 mol.) ofN-(N-tert.-butyloxycarbonylaminoxy-acetyl)-aminoxy-acetic acid hydrazideare dissolved in 25 m1. of abs. dimethylformamide. 3.07 g. (0.07 mol.)of N-tert.-butyloxycarbonyl-aminoxy-acetic acid pentachlorophenyl esterare added to the reacton mixture, which is stirred for one hour at roomtemperature and then left to stand for 16 hours. The solvent is thendistilled off under reduced pressure, in a water bath of max. 50 C., theresidue is dissolved in 3-0 ml. of ethyl acetate, and the solutionextracted three times with 7 ml. of 0.1 N hydrochloric acid solution andsubsequently twice with 7 ml. of water. The organic layer is dried overanhydrous sodium sulphate, and the solvent is distilled off. 20 ml. of a4 mol./ liter solution of hydrogen chloride in ethyl acetate are addedto the residue, the mixture is stirred for 15 minutes at roomtemperature; 60 ml. of abs. diethyl ether are then added, theprecipitated crystals are collected, washed with diethyl ether, driedover phosphorus pentoxide, and then recrystallized from ethanol/ ether.1.70 g. of N'- (N-aminoxy-acetyl)-aminoxy-acetyl-N-aminoxy-acetyl-hydrazine dihydrochloride (75%) are obtained; M.P.176178 C.; R;=0.18.

Analysis.Calculated (percent): C, 22.2; H, 4.8; N, 21.6; C1, 21.9. Found(percent): C, 22.2; H, 4.9; N, 21.4; C1, 22.0.

EXAMPLE 10 (a) N-(N"-tert.-butyloxycarbonyl-DL or aminoxy propicinyl) Nisonicotinoyl-hydrazine pentachloropheno ate 1.73 g. (0.0038 mol.) ofDL-a-tert.-butyloxycarbonylaminoxy-propionic acid pentachlorophenylester are dissolved in 15.0 ml. of abs. dimethylformamide under stirringat room temperature. 0.57 g. (0.00415 mol.) of iso nicotinic acidhydrazide are then added, and the reaction mixture is left to stand for16 hours. The solvent is then distilled off under reduced pressure at atemperature of max. 50 C., and the residue is recrystallized from ethylacetate. 2.00 g. (91%) of N-(N"-tert.-butyloxycarbonyl-Dl-a-aminoxy-propionyl)-N'-isonicotinoyl-hydrazine pentachlorophenolateare obtained; M.P. 136-138 C.; R;=0.22.

Analysis.Calculated (percent): C, 40.7; H, 3.4; CI, 30.1. Found(percent): C, 40.7; H, 3.5; CI, 30.1.

(b) N-DL-a-aminoxy-propiony1-N'-isonicotinoylhydrazine dihydrobromide 9EXAMPLE 11 (a) N- (N"-tert.-butyloxycarbonyl-tx-aminoxy-propionyl) N'-(N"'-benzyloxycarbonyl-glycyl) -hydrazine 0.96 g. (0.0043 mol.) ofN-benzyloxycarbonyl-glycylhydrazine are dissolved in 20 ml. of abs.dimethylformamide under stirring at room temperature. 1.82 g. (0.0040mol.) of N-tert.-butyloxycarbonyl-aminoxy-propionic acidpentachlorophenyl ester are added to the solution and the mixture isleft to stand for 16 hours. The solvent is then distilled ofi underreduced pressure, the residue is dissolved in 25 ml. of ethyl acetate,the solution washed three times with ml. of N hydrochloric acid, andwith 5 ml. of water, the organic layer is dried over anhydrous sodiumsulphate, and the solvent is distilled off under reduced pressure. Theresidue is dissolved in hot ethyl acetate and n-heptane is added untilthe crystallization of the product is just beginning. After cooling, theprecipitated crystals are collected. 1.21 g. (74%) of N (N"tert.-butyloxycarbonyl-a-aminoxy-propionyl)- N (N"'benzyloxycarbonyl-glycyl)-hydrazine are obtained; M.P. 99l00 C.; [a]=+5l (c.=l, ethanol); R;=0.27.

Analysis.Calculated (percent): Found (percent): C, 52.7; H, 6.5.

(b) N -a-aminoxy-propionyl-N'-glycyl-hydrazine dihydrobromide 1.10 g.(0.0027 mol.) of N-(N"-tert.-butyloxy-carbonylaaminoxy-propionyl)-N'-(N'-benzyloxycarbonyl glycyl)-hydrazine aretreated with hydrobromic acid dissolved in glacial acetic acid, asdescribed in Example 1c). 0.66 g. (91%) ofN-a-aminoxy-propionyl-N'-glycyl-hydrazine dihydrobromide are obtained;M.P. 170-175 C., [a] =+30 (c.=0.8, ethanol).

Analysis.Calculated (percent): C, 17.8; H, 4.2; Br, 47.3. Found(percent): C, 1.77; H, 4.3; Br, 47.5.

EXAMPLE 12 (a) N-(N"-tert.-butyloxycarbonyl-u-aminoxy-B-phenylpropionyl)-N-isonicotinoyl-hydrazine 3.28 g. (0.0062 mol.) ofN-tert.-butyloxycarbonyl-aaminoxy-B-phenyl-propionic acidpentachlorophenyl ester are dissolved in 20 ml. of abs.dimethylformamide, 0.98 g. (0.0065 mol.) of isonicotinic acid hydrazideare added to the solution and the mixture is left to stand for 16 hours.The solvent is then distilled 01f under reduced pressure in a water bathof max. 50 C. The residue is dissolved in 25 ml. of ethyl acetate andthe ethyl acetate solution is extracted three times with ml. of aqueousN hydrochloric acid. The aqueous extracts are unified, and thenneutralized by the addition of solid sodium bicarbonate. Theprecipitated crystals are collected, washed with water and dried, 2.35g. (95%) of N-(N"-tert.-butyloxycarbonyl a aminoxy Bphenyl-propionyl)-N'-isonicotinoyl-hydrazin are obtained; M.P. 98-102"C.; [a] =67 (c.=1, ethanol); R =0.24.

Analysis.-Ca1culated (percent): C, 60.0; H, 6.0; N, 14.0. Found(percent): C, 59.9; H, 6.1; N, 13.9.

(b) N-a-aminoxy-B-phenyl-propionyl-N-isonicotinoylhydrazinedihydrobromide 2.00 g. (0.005 mol.) of N-(N"-tert.-butyloxycarbony1-u-aminoxy B phenyl-propionyl)-N-isonicotinoyl hydrazine are treated with20 ml. of a 4 moL/Iiter solution of hydrobromic acid in glacial aceticacid, as described in Example 1(c). 2.29 g. (99%) ofN-a-aminoxy-fl-phenyl- 10 propionyl-N-isonicotinoyl-hydrazinedihydrobromid are obtained; M.P. 168-170 C.; [a] =-28 (c.=1, ethanol).

Analysis.-Calculated (percent): C, 39.0; H, 3.9; Br, 34.6. Found(percent): C, 38.9; H, 4.0; Br, 34.6.

EXAMPLE 13 N-(N"-acetyl-DL-ot-aminoxy-fl-phenyl-propionyl)- N'-isonicotinoyl-hydrazine 3.25 g. (0.0087 mol.) ofN-(DL-a-aminoxy-fi-phenylpropionyl) N isonicotinoyl-hydrazinedihydrochloride are dissolved in 20 ml. abs. pyridine. The solution iscooled to 5 C., 2.40 ml. (0.0174 mol.) of triethylarnine are addedthereto and then 1.24 ml. (0.0174 mol.) of acetyl chloride are addeddropwise, under further cooling to the mixture. The reaction is thenallowed to warm up to room temperature and stirred for further 20minutes. 1.20 ml. (0.0087 mol.) of triethylamine are added to themixture and the stirring is continued for a further hour. The mixture isdiluted with ml. of water and extracted three times with ethyl acetate(25 ml. each). The unified ethyl acetate extract is washed with 25 ml.of water, dried over anhydrous sodium sulphate and evaporated to drynessunder reduced pressure. The residue is recrystallized from hot ethylacetate. 1.98 g. (66.5%) of N-(N-acetyl-DL-a-aminoxy-;S'-phenyl-propionyl) N isonicotinoylhydrazine areobtained; M.P. 173-174 C.; R =0.43.

Analysis.Calculated (percent): C, 59.6; H, 5.3; N, 16.4. Found(percent): C, 59.7; H, 5.3; N, 16.3.

EXAMPLE 14 N-(N"-benzoyl-aminoxyacetyl)-N'-isonicotinoylhydrazine 7.44g. (0.20 mol.) of N aminoxyacetyl-N'-isonicotionoyl-hydrazinedihydrobromide are dissolved in 25 ml. of Water and 1.60 g. (0.040 mol.)of solid sodium hydroxide are added under stirring to the solutioncooled previously to 5 C. 2.52 g. (0.030 mol.) of solid sodiumbicarbonate are then added to the reaction mixture and 3.50 ml. (0.030mol.) of benzoyl chloride are added dropwise thereto in 30 minutes,while the temperature of the reaction mixture is held at 5 C. Themixture is stirred for further 2 hours at room temperature, theprecipitated product is then collected and recrystallized (withoutprevious drying) from water. 5.42 g. (79%) of N-(N"-benzoyl-aminoxyacetyl-N'-isonicotinoyl-hydrazine are obtained; M.P.-156" C.; R =0.35.

Analysis.Calculated (percent): C, 55.8; H, 4.7; N, 16.3. Found(percent): C, 55.7; H, 4.7; N, 16.2.

What we claim is:

1. N-aminoxy-acetyl N isonicotinoyl-hydrazine and the pharmaceuticallyacceptable acid addition salts thereof.

OTHER REFERENCES Burger: Medicinal Chemistry, Third edition, Part I,Wiley-Interscience, pp. 414-415 (1970).

ALAN L. ROTMAN, Primary Examiner U.S. Cl. X.R.

